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Particle Display: A Quantitative Screening Method for Generating High‐Affinity Aptamers
Author(s) -
Wang Jinpeng,
Gong Qiang,
Maheshwari Nupur,
Eisenstein Michael,
Arcila Mary Luz,
Kosik Kenneth S.,
Soh H. Tom
Publication year - 2014
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201309334
Subject(s) - aptamer , sorting , selection (genetic algorithm) , cell sorting , dna , particle (ecology) , nanotechnology , chemistry , computational biology , particle size , computer science , biological system , combinatorial chemistry , biophysics , biology , materials science , cell , microbiology and biotechnology , artificial intelligence , biochemistry , algorithm , ecology
We report an aptamer discovery technology that reproducibly yields higher affinity aptamers in fewer rounds compared to conventional selection. Our method (termed particle display) transforms libraries of solution‐phase aptamers into “aptamer particles”, each displaying many copies of a single sequence on its surface. We then use fluorescence‐activated cell sorting (FACS) to individually measure the relative affinities of >10 8 aptamer particles and sort them in a high‐throughput manner. Through mathematical analysis, we identified experimental parameters that enable optimal screening, and demonstrate enrichment performance that exceeds the theoretical maximum achievable with conventional selection by many orders of magnitude. We used particle display to obtain high‐affinity DNA aptamers for four different protein targets in three rounds, including proteins for which previous DNA aptamer selection efforts have been unsuccessful. We believe particle display offers an extraordinarily efficient mechanism for generating high‐quality aptamers in a rapid and economic manner, towards accelerated exploration of the human proteome.